NUCLEASE-RESISTANT DNA ANALOGUES
The present invention provides stable, nuclease-resistant TNA, TNA-DNA and TNA-RNA oligonucleotides, wherein the oligonucleotides are completely resistant to enzymatic degradation for at least 24-72 hours. Methods of synthesis and use in diagnostic and therapeutic applications are also provided.
1 . A stable, nuclease-resistant TNA-RNA oligonucleotide, wherein the TNA-RNA oligonucleotide comprises an effective amount of TNA to provide resistance to enzymatic degradation in a biological environment wherein at least half the oligonucleotides are TNA.
2 . The nuclease-resistant TNA-RNA oligonucleotide of claim 1 , wherein the TNA-RNA oligonucleotide is resistant to enzymatic degradation by snake venom phosphodiesterase, RNAse A, RQ1 DNAse, and Turbo DNAse for at least 72 hours.
3 . The nuclease-resistant TNA-RNA oligonucleotide of claim 1 , wherein the effective amount of TNA comprises at least two TNA nucleic acids.
4 . The nuclease-resistant TNA-RNA oligonucleotide of claim 1 , wherein the effective amount of TNA comprises at least three TNA nucleic acids.
5 . The nuclease-resistant TNA-RNA oligonucleotide of claim 1 , wherein the TNA residues alternate with RNA residues.
10 . The nuclease-resistant TNA-RNA oligonucleotide of claim 1 , wherein the TNA-RNA oligonucleotide forms a duplex wherein a portion of the oligonucleotide comprises TNA that is able to anneal to a complementary portion of the oligonucleotide containing RNA, wherein the duplex is resistant to enzymatic degradation in a biological environment of the TNA-RNA duplex.
11 . The nuclease-resistant TNA-RNA oligonucleotide of claim 10 , wherein the TNA-RNA duplex is resistant to enzymatic degradation by snake venom phosphodiesterase, RNAse A, RQ1 DNAse, and Turbo DNAse for at least 24 hours.
12 . The nuclease-resistant TNA-RNA oligonucleotide of claim 10 , wherein the TNA-RNA duplex is resistant to enzymatic degradation for at least 72 hours.
13 . A TNA-RNA duplex comprising a RNA strand and a complementary TNA strand, wherein the duplex comprises an effective amount of TNA to provide resistance to enzymatic degradation in a biological environment and wherein the TNA strand is able to anneal to the RNA strand.
14 . The TNA-RNA duplex of claim 13 , wherein the duplex is resistant to enzymatic degradation for at least 24 hours.
15 . The TNA-RNA duplex of claim 14 , wherein the duplex is resistant to enzymatic degradation for at least 72 hours.
16 . A method of preparing a nuclease-resistant TNA-RNA oligonucleotide of claim 1 , the method comprising inserting an effective amount of TNA into a sample of RNA to yield a TNA-RNA oligonucleotide, wherein the TNA-RNA oligonucleotide is resistant to enzymatic degradation in a biological environment.
17 . The method of claim 16 , wherein the TNA-RNA oligonucleotide is resistant to enzyme degradation for at least 24 hours.
18 . The method of claim 16 , wherein the TNA residues alternate with RNA residues.
19 . The method of claim 16 , wherein the TNA-RNA oligonucleotide is resistant to enzyme degradation for at least 72 hours.
20 . The method of claim 16 , wherein the TNA-RNA oligonucleotide comprises a portion of RNA continuous with a portion of the oligonucleotide comprising TNA that is able to anneal to a complementary portion of the oligonucleotide containing RNA.